Label-Free Quantitative Proteomic Study of The Effect of Dihydrotestosterone (DHT) On Rat Ovarian GCs

Dihydrotestosterone (DHT) is a main androgen in the human body. Previous reports have shown that DHT can affect the proliferation, apoptosis and estrogen and progesterone secretion of ovarian granulosa cells (GCs). An imbalance in DHT secretion leads to GC dysfunction and follicular development disorder.Therefore, exploring the influence of DHT on GCs is necessary. The purpose of this study was to analyze the effect of DHT on GCs through label-free quantitative proteomics (LFQP). After primary cultured rat GCs were treated with DHT (10-8 mol/L), the effect of DHT on GCs was analyzed by LFQP, and some of the differentially expressed proteins (DEPs) were verified by western blotting.A total of 6124.0 proteins were identified, of which 4496.0 were quantifiable. Compared with the control group, 28 proteins were upregulated and 10 were downregulated after DHT intervention. The subcellular localization of DEPs indicates that DHT is involved in the proliferation, migration, molding and metabolism of GCs. Gene Ontology (GO) revealed that DHT downregulated the oxygen transport capacity and oxygen-binding protein of GCs. Orthologous Groups of proteins (COG/KOG) showed that DHT had an important effect on the survival, growth and apoptosis of GCs. Kyoto Encyclopedia of Genes and Genomes (KEGG) revealed that DHT promotes metabolism, amino acid degradation, chemical carcinogenesis, platelet activation and vasoconstriction in GCs. The western blot results were consistent with the proteomics results. Mark3 and Mre11a are DEPs that were upregulated, and Fth1 and Nqo1 were downregulated, which indicated that DHT could promote the proliferation of GCs.This study comprehensively analyzes the impact of DHT on GCs through LFQP and provides clues for further research.

Specialization in a nitrogen-fixing symbiosis: proteome differences between Sinorhizobium medicae bacteria and bacteroids

Using tandem mass spectrometry (MS/MS), we analyzed the proteome of Sinorhizobium medicae WSM419 growing as free-living cells and in symbiosis with Medicago truncatula. 3215 proteins were identified, over half of the ORFs predicted from the genomic sequence. The abundance of 1361 proteins displayed strong lifestyle bias. 1131 proteins had similar levels in bacteroids and free-living cells, and the low levels of 723 proteins prevented statistically significant assignments. Nitrogenase subunits comprised ~12% of quantified bacteroid proteins. Other major bacteroid proteins included symbiosis-specific cytochromes and FixABCX, which transfer electrons to nitrogenase. Bacteroids had normal levels of proteins involved in amino acid biosynthesis, glycolysis/gluconeogenesis and the pentose phosphate pathway, but several amino acid degradation pathways were repressed. This suggests bacteroids maintain a relatively independent anabolic metabolism. TCA cycle proteins were highly expressed in bacteroids and no other catabolic pathway emerged as an obvious candidate to supply energy and reductant to nitrogen fixation. Bacterial stress response proteins were induced in bacteroids. Many WSM419 proteins that are not encoded in Sinorhizobium meliloti Rm1021 were detected and understanding the functions of these proteins might clarify why S. medicae WSM419 forms a more effective symbiosis with M. truncatula than S. meliloti Rm1021.

Network Analysis of the Effects of Long Non-coding RNAs in Artemisinin treatment of Atherosclerosis in APOE-/- Mice

IntroductionAtherosclerosis has become a worldwide medical burden. Our previous studies have shown that Artemisinin(ART) had the capability to reduce atherosclerosis. Emerging evidence indicates that Long non-coding RNAs(lncRNAs) are engaged in the formation of atherosclerosis. However, whether lncRNAs might participate in the mechanism through which artemisinin mitigates atherosclerosis has not been reported.Material and methodsEight-week-old apolipoprotein E deficient (APOE-/-) mice were divided into two groups, one of which was treated with Artemisinin. Red oil O staining was used to measure the sizes of Atherosclerotic lesion. We conducted deep sequencing to investigate lncRNA profiles in the aorta tissue in high-fat diet fed APOE knockdown mice with and without artemisinin treatment. CeRNA network, Kyoto Encyclopedia of Genes and Genomes(KEGG) and Gene Ontology(GO) analyses were constructed through bioinformatics analysis. RT-PCR was used to validate the differentially expressed lncRNAs.ResultsA total of 102 lncRNAs and 4,630 mRNAs were differentially expressed (p<0.05) between the Artemisinin treatment group and atherosclerosis model group. KEGG and GO analyses indicated that the categories metabolic process, specific amino acid degradation and PI3K-Akt signaling pathway are involved in the effects of artemisinin treatment in atherosclerosis(qvalue<0.05). LncRNA ENSMUST00000099676.4, ENSMUST00000143673.1, ENSMUST00000070085.5 and ENSMUST00000224554 might be engaged in treatment mechanism through which Artemisinin alleviates Atherosclerosis.ConclusionsThese findings indicated the possible mechanism and therapeutic role of lncRNAs in Artemisinin treatment of atherosclerosis and provided a theoretical basis for the future application of Artemisinin in patients with atherosclerosis.

Effects of Multi-Species Direct-Fed Microbial Products on Ruminal Metatranscriptome and Carboxyl-Metabolome of Beef Steers

We examined the effects of two direct-fed microbial (DFM) products containing multiple microbial species and their fermentation products on ruminal metatranscriptome and carboxyl-metabolome of beef steers. Nine ruminally-cannulated Holstein steers were assigned to 3 treatments arranged in a 3 × 3 Latin square design with three 21-d periods. Dietary treatments were (1) Control (CON; basal diet without additive), (2) Commence (PROB; basal diet plus 19 g/d of Commence), and (3) RX3 (SYNB; basal diet plus 28 g/d of RX3). Commence and RX3 are both S. cerevisiae-based DFM products containing several microbial species and their fermentation products. Mixed ruminal contents collected multiple times after feeding on day 21 were used for metatranscriptome and carboxyl-metabolome analysis. Partial least squares discriminant analysis revealed a distinct transcriptionally active taxonomy profiles between CON and each of the PROB and SYNB samples. Compared to CON, the steers fed supplemental PROB had 3 differential (LDA ≥ 2.0; p ≤ 0.05) transcriptionally active taxa, none of which were at the species level, and those fed SYNB had eight differential (LDA > 2.0, p ≤ 0.05) transcriptionally active taxa, but there was no difference (p > 0.05) between PROB and SYNB. No functional microbial genes were differentially expressed among the treatments. Compared with CON, 3 metabolites (hydroxylpropionic acid and 2 isomers of propionic acid) were increased (FC ≥ 1.2, FDR ≤ 0.05), whereas 15 metabolites, including succinic acid and fatty acid peroxidation and amino acid degradation products were reduced (FC ≤ 0.83, FDR ≤ 0.05) by supplemental PROB. Compared with CON, 2 metabolites (2 isomers of propionic acid) were increased (FC ≥ 1.2, FDR ≤ 0.05), whereas 2 metabolites (succinic acid and pimelate) were reduced (FC ≤ 0.83, FDR ≤ 0.05) by supplemental SYNB. Compared to SYNB, supplemental PROB reduced (FC ≤ 0.83, FDR ≤ 0.05) the relative abundance of four fatty acid peroxidation products in the rumen. This study demonstrated that dietary supplementation with either PROB or SYNB altered the ruminal fermentation pattern. In addition, supplemental PROB reduced concentrations of metabolic products of fatty acid peroxidation and amino acid degradation. Future studies are needed to evaluate the significance of these alterations to ruminal fatty acid and amino acid metabolisms, and their influence on beef cattle performance.

The proteomic characteristics of airway mucus from critical ill COVID-19 patients

Background: The pandemic of the coronavirus disease 2019 (COVID-19) has brought a global public health crisis. However, the pathogenesis underlying COVID-19 are barely understood. Methods: In this study, we performed proteomic analyses of airway mucus obtained by bronchoscopy from severe COVID-19 patients. In total, 2351 and 2073 proteins were identified and quantified in COVID-19 patients and healthy controls, respectively. Results: Among them, 92 differentiated expressed proteins (DEPs) (46 up-regulated and 46 down-regulated) were found with a fold change > 1.5 or < 0.67 and a p-value < 0.05, and 375 proteins were uniquely present in airway mucus from COVID-19 patients. Pathway and network enrichment analyses revealed that the 92 DEPs were mostly associated with metabolic, complement and coagulation cascades, lysosome, and cholesterol metabolism pathways, and the 375 COVID-19 only proteins were mainly enriched in amino acid degradation (Valine, Leucine and Isoleucine degradation), amino acid metabolism (beta-Alanine, Tryptophan, Cysteine and Methionine metabolism), oxidative phosphorylation, phagosome, and cholesterol metabolism pathways. Conclusions: This study aims to provide fundamental data for elucidating proteomic changes of COVID-19, which may implicate further investigation of molecular targets directing at specific therapy.

Iron is not everything: unexpected complex metabolic responses between iron-cycling microorganisms

Coexistence of microaerophilic Fe(II)-oxidizers and anaerobic Fe(III)-reducers in environments with fluctuating redox conditions is a prime example of mutualism, in which both partners benefit from the sustained Fe-pool. Consequently, the Fe-cycling machineries (i.e., metal-reducing or –oxidizing pathways) should be most affected during co-cultivation. However, contrasting growth requirements impeded systematic elucidation of their interactions. To disentangle underlying interaction mechanisms, we established a suboxic co-culture system of Sideroxydans sp. CL21 and Shewanella oneidensis. We showed that addition of the partner’s cell-free supernatant enhanced both growth and Fe(II)-oxidizing or Fe(III)-reducing activity of each partner. Metabolites of the exometabolome of Sideroxydans sp. CL21 are generally upregulated if stimulated with the partner´s spent medium, while S. oneidensis exhibits a mixed metabolic response in accordance with a balanced response to the partner. Surprisingly, RNA-seq analysis revealed genes involved in Fe-cycling were not differentially expressed during co-cultivation. Instead, the most differentially upregulated genes included those encoding for biopolymer production, lipoprotein transport, putrescine biosynthesis, and amino acid degradation suggesting a regulated inter-species biofilm formation. Furthermore, the upregulation of hydrogenases in Sideroxydans sp. CL21 points to competition for H2 as electron donor. Our findings reveal that a complex metabolic and transcriptomic response, but not accelerated formation of Fe-end products, drive interactions of Fe-cycling microorganisms.

Effect of wet/dry, fresh liquid, fermented whole diet liquid, and fermented cereal liquid feeding on feed microbial quality and growth in grow-finisher pigs

Fermented liquid feeding has proved beneficial for weaner pigs; however, there is limited research on its effect on the growth and feed conversion efficiency (FCE) of grow-finisher pigs. Microbial decarboxylation of amino acids is associated with whole diet fermentation, while wet/dry and liquid feeding reportedly improve growth compared with dry feeding. The objective of this study was to determine the effect of wet/dry feeding and fresh, fermented whole diet, and fermented cereal liquid feeding on pig growth, feed efficiency, and carcass quality in grow-finisher pigs. Pigs were allocated to one of four dietary treatments in two experiments: 1) Single-space wet/dry feeders (WET/DRY), 2) Fresh liquid feeding (FRESH), 3) Fermented cereal liquid feeding where the cereal fraction (38% barley, 40% wheat) of the diet was fermented prior to feeding (FERM-CER), and 4) Fermented whole diet liquid feeding where the whole diet was fermented prior to feeding (FERM-WH). In exp. 1, pigs were fed the experimental diets for 68 d prior to slaughter (29.8 kg ± 0.92 SE to 102.3 kg ± 0.76 SE). Overall, average daily gain (ADG) was 1,094, 1,088, 1,110, and 955 g/d (SE = 13.0; P &lt; 0.001) and FCE was 2.26, 2.37, 2.40, and 2.88 (SE = 0.031; P &lt; 0.001) for treatments one through four, respectively. Pigs fed FERM-WH were lighter at slaughter than pigs fed the other three treatments (P &lt; 0.001). In exp. 2, pigs were on treatment for 26 d prior to slaughter (85.3 kg ± 1.69 SE to 117.5 kg ± 0.72 SE). Overall, ADG in exp. 2 was 1,103, 1,217, 1,284, and 1,140 g/d (SE = 27.9; P &lt; 0.01) and FCE was 2.78, 2.99, 2.95, and 3.09 g/g (SE = 0.071; P = 0.05), for treatments one through four, respectively. There were no significant differences observed between treatments for apparent total tract digestibility of dry matter, organic matter, nitrogen, gross energy, or ash. Higher lactic acid bacteria counts and lower Enterobacteriaceae counts and pH were observed in FERM-CER and FERM-WH compared with WET/DRY and FRESH. Ethanol concentrations were almost 4-fold higher in FERM-CER troughs than FRESH troughs and 5-fold higher in FERM-WH than FRESH troughs. To conclude, FERM-WH resulted in poorer growth and FCE compared with WET/DRY, FRESH, and FERM-CER, probably due to amino acid degradation and a loss in gross energy found in FERM-WH.

Prenylquinones in Human Parasitic Protozoa: Biosynthesis, Physiological Functions, and Potential as Chemotherapeutic Targets

Human parasitic protozoa cause a large number of diseases worldwide and, for some of these diseases, there are no effective treatments to date, and drug resistance has been observed. For these reasons, the discovery of new etiological treatments is necessary. In this sense, parasitic metabolic pathways that are absent in vertebrate hosts would be interesting research candidates for the identification of new drug targets. Most likely due to the protozoa variability, uncertain phylogenetic origin, endosymbiotic events, and evolutionary pressure for adaptation to adverse environments, a surprising variety of prenylquinones can be found within these organisms. These compounds are involved in essential metabolic reactions in organisms, for example, prevention of lipoperoxidation, participation in the mitochondrial respiratory chain or as enzymatic cofactors. This review will describe several prenylquinones that have been previously characterized in human pathogenic protozoa. Among all existing prenylquinones, this review is focused on ubiquinone, menaquinone, tocopherols, chlorobiumquinone, and thermoplasmaquinone. This review will also discuss the biosynthesis of prenylquinones, starting from the isoprenic side chains to the aromatic head group precursors. The isoprenic side chain biosynthesis maybe come from mevalonate or non-mevalonate pathways as well as leucine dependent pathways for isoprenoid biosynthesis. Finally, the isoprenic chains elongation and prenylquinone aromatic precursors origins from amino acid degradation or the shikimate pathway is reviewed. The phylogenetic distribution and what is known about the biological functions of these compounds among species will be described, as will the therapeutic strategies associated with prenylquinone metabolism in protozoan parasites.

Aromatic Profile, Physicochemical and Sensory Traits of Dry-Fermented Sausages Produced without Nitrites Using Pork from Krškopolje Pig Reared in Organic and Conventional Husbandry

Dry-fermented sausages were produced in a traditional way, without addition of nitrites and starter cultures, from meat of an autochthonous breed (Krškopolje pig) raised either in a conventional indoor or organic husbandry system. Physicochemical and sensory analyses were performed at the end of processing to characterize their quality. Dry-fermented sausages from organic pork retained more moisture, which resulted in higher water activity and softer texture (instrumental and sensory). They were more oxidized (higher thiobarbituric acid reactive substances (TBARS)), in agreement with more unsaturated fatty acid profile, a higher score for rancid taste, and a higher relative abundance of volatiles from lipid β-oxidation. Overall, dry-fermented sausages from organic pork had lower levels of volatile compounds, particularly, those originating from spices (despite the same quantity added) and lower levels of amino-acid degradation. Sensory analysis showed that dry-fermented sausages from organic pork had less intensive and vivid color, tasted more bitter and sour, and had more off-tastes. The observed differences could be related to initial differences in raw material (differences in meat pH and level of polyunsaturated fatty acids) affecting the process of fermentation.